关键词：
腹泻;家兔;复方救必应;蛋白质-蛋白质相互作用;网络药理学
摘 要：
为了深入研究复方救必应(由救必应、香附、东风桔组成)对腹泻家兔的治疗效果和作用机制,试验将 54 只1 月龄健康试验兔随机分为空白组、模型组和试验组(每组 18 只),空白组正常饲喂兔饲料,模型组和试验组家兔通过高脂高糖饮食+生物因子法构建腹泻模型,模型组家兔饲喂高脂饲料,试验组家兔饲喂按照每只家兔 2.8 g/d的剂量用复方救必应拌料的高脂饲料,试验期为14 d.采集家兔耳缘静脉血,用相应的酶联免疫吸附测定试剂盒检测血清白细胞介素-1(IL-1)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量;利用ETCM获得救必应、香附、东风桔3 味中药的活性成分及作用靶点,利用Genecard获得与腹泻疾病相关的基因,进一步获得复方救必应治疗腹泻的潜在靶点;利用Metascape对潜在靶点进行GO功能注释和KEGG信号通路分析;利用Cytoscape 3.7.2 软件构建"中药-成分-靶点-通路-疾病"拓扑网络并分析蛋白质-蛋白质相互作用(PPI)网络.结果表明:与空白组比较,模型组家兔腹泻率和死亡率明显升高;与模型组比较,试验组家兔腹泻率和死亡率明显降低;试验组家兔血清中IL-1、IL-6、TNF-α含量均极显著低于模型组(P<0.01);3 味中药共检索到 17 种活性成分,127 个治疗腹泻的潜在靶点,这些靶点主要被注释到细胞黏附调节、炎症应答、白细胞迁徙等生物过程,以及PI3K-Akt信号通路、趋化因子信号通路和丝裂原活化蛋白激酶(MAPK)信号通路中;β-莎草醇、香附醇酮、香附醇、β-香树脂醇、isocyperol、鼠李素-3-O-鼠李糖基(1-4)-吡喃鼠李糖甙在复方救必应发挥功能过程中可能具有重要作用,是主要的活性成分;在PPI网络中,IL-6、AKT1、TNF、ACTB为关键节点.说明复方救必应具有一定的治疗家兔腹泻的作用,其作用机制涉及多成分-多靶点-多通路.
译 名：
Study on the therapeutic effects and mechanism of compound Jiu bi ying on rabbit diarrhea
关键词：
diarrhea%rabbit%compound jiu bi ying%protein-protein interaction%network pharmacology
摘 要：
In order to deeply investigate the therapeutic effects and mechanism of compound Jiu bi ying(consisting of Ilex rotunda Thunb,Cyperus rotundus L.Atalantia buxifolia[Poir.]Oliv in equal amounts)on diarrhea in rabbits,54 one-month-old healthy rabbits were randomly divided into blank group,model group,and experimental group,with 18 rabbits in each group.The blank group was fed normally,while the model group and the test group were constructed by feeding rabbits with high-fat and high-sugar diet + biological factors to establish a diarrhea model.The experimental group of rabbits was treated with compound Jiu bi ying mixed with feed at a dose of 2.8 g/d for 14 days.The serum levels of interleukin-1(IL-1),interleukin-6(IL-1)and tumor necrosis factor-α(TNF-α)were detected by ELISA kit.The active ingredients and application points of the three Chinese herbs jiu bi ying,Xiangfu and Dongfeng Tangerine were obtained by using ETCM,and the genes related to diarrhea disease were obtained by using Genecard to further obtain the potential targets of compound Jiubiying in the treatment of diarrhea.GO function annotation and KEGG signaling pathway analysis of potential targets were performed by using Metascape.The"TCM-Component-Target-pathway-disease"topological network was constructed and protein-protein interaction(PPI)network was analyzed by using Cytoscape 3.7.2 software.The results showed that compared with the blank group,the diarrhea rate and mortality of rabbits in the model group were significantly increased.Compared with model group,the diarrhea rate and mortality of rabbits in experimental group were significantly reduced.The contents of IL-1,IL-6 and TNF-α in serum of experimental group were significantly lower than those of model group(P<0.01).A total of 17 active ingredients and 127 potential targets for the treatment of diarrhea were detected from the three Chinese herbs.These targets were mainly annotated in biological processes such as regulation of cell adhesion,inflammatory response,and leukocyte migration,as well as in the PI3K-Akt signaling pathway,chemokine signaling pathway and mitogen-activated protein kinase(MAPK)signaling pathway.β-peryol,vanillone,vanillol,β-vanillinol,isocyperol,and rhamnose-3-O-rhamnosyl(1-4)-rhamnoside might play an important role in the functional process of compound Jiu bi ying,and were the main active ingredients.In the PPI network,IL-6,AKT1,TNF and ACTB were the key nodes.These results indicated that compound Jiu bi ying had a certain effect on the treatment of rabbit diarrhea,and its mechanism involved multi-component-multi-target-multi-pathway.
